Pattern stability under cell culture conditions—A comparative study of patterning methods based on PLL-g-PEG background passivation

Lussi, Jost W.; Falconnet, Didier; Hubbell, Jeffrey A.; Textor, Marcus; Csúcs, Gábor

doi:10.1016/j.biomaterials.2005.11.027

Cited by 94 publications

(84 citation statements)

References 41 publications

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“…No cells were found in the passivated region, confirming the results of earlier studies that cells can not attach, spread, or move on PLL-g-PEG treated surfaces [Csucs et al, 2003;Lussi et al, 2006]. Cells crawling on the Fnprinted region turned around as soon as they reached the boundary between printed and PLL-g-PEG treated surface, providing further evidence that unprinted and PLLg-PEG passivated substrates were cell-repulsive and non-adhesive.…”

Section: Migration Behaviorsupporting

confidence: 91%

Locomotion of fish epidermal keratocytes on spatially selective adhesion patterns

Csúcs

Quirin

Danuser

2007

Cell Motil. Cytoskeleton

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Cell migration results from forces generated by assembly, contraction, and adhesion of the cytoskeleton. To address how these forces integrate in space and time, novel assays are required that allow spatial separation of the different force categories. We used micro-contact printing of fibronectin on glass substrates to study the effect of adhesion patterns on fish epidermal keratocytes locomotion. Cells migrated at similar speeds on homogeneously adhesive substrates and on patterns with 5 microm-wide adhesive stripes interleaved by non-adhesive stripes with a width varied between 5 and 13 microm. The leading edge protruded on adhesive stripes and lagged behind on non-adhesive stripes. On patterns with non-adhesive stripes wider than 13 microm cells halted, although the lamellipodium did not collapse. High correlation was found between the widths of protruding and lagging edge segments and the widths of the underlying stripes. We explain our data by the force balances between actin polymerization, contraction and adhesion on fibronectin stripes; and between actin polymerization, contraction and lamellipodium-internal elastic tension on non-adhesive stripes. We tested our model further by blocking lamellipodium actin network contraction and polymerization. In both experiments we observed that cells eventually lost their ability to move. However, the two perturbations induced distinct morphological responses. The data suggested that forces powering forward motion of keratocytes are largely associated with network assembly whereas contraction maintains cell polarity. This study establishes spatially selective adhesion substrates and cell morphological readouts as a means to elucidate the mechanical balance between substrate adhesion and cytoskeleton-internal tension in cell migration.

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Section: Migration Behaviorsupporting

confidence: 91%

Locomotion of fish epidermal keratocytes on spatially selective adhesion patterns

Csúcs

Quirin

Danuser

2007

Cell Motil. Cytoskeleton

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“…Microcontacting printing (mCP) is superior to selective molecular assembly patterning (SMAP) and molecular assembly patterning by li-off (MAPL) methods 40 and is by far the most popular technique for biologists to pattern proteins and cells. mCP can also be extended to the patterned transfer of PL or PO.…”

Section: Microcontact Printing Polylysine On Peg Adlayersmentioning

confidence: 99%

Micropatterning neuronal networks

Hardelauf

Waide

Sisnaiske

et al. 2014

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Spatially organised neuronal networks have wide reaching applications, including fundamental research, toxicology testing, pharmaceutical screening and the realisation of neuronal implant interfaces. Despite the large number of methods catalogued in the literature there remains the need to identify a method that delivers high pattern compliance, long-term stability and is widely accessible to neuroscientists. In this comparative study, aminated (polylysine/polyornithine and aminosilanes) and cytophobic (poly(ethylene glycol) (PEG) and methylated) material contrasts were evaluated. Backfilling plasma stencilled PEGylated substrates with polylysine does not produce good material contrasts, whereas polylysine patterned on methylated substrates becomes mobilised by agents in the cell culture media which results in rapid pattern decay. Aminosilanes, polylysine substitutes, are prone to hydrolysis and the chemistries prove challenging to master. Instead, the stable coupling between polylysine and PLL-g-PEG can be exploited: Microcontact printing polylysine onto a PLL-g-PEG coated glass substrate provides a simple means to produce microstructured networks of primary neurons that have superior pattern compliance during long term (>1 month) culture.

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“…This circumvented the issue of impaired fidelity of protein patterns as well as poor long-term stability that was reported for the PLL-g-PEG coating when used in 2D protein pattern applications. 36 The fidelity of the microwell functionalization was maintained for up to 7 days in the culture of human bone marrow-derived mesenchymal progenitor cells, as demonstrated by the improved confinement of the cells within just the wells and not the plateau.…”

Section: Engineering Microwells As 2d Single Cell Microenvironmentsmentioning

confidence: 99%

Integration column: microwell arrays for mammalian cell culture

et al. 2009

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Pattern stability under cell culture conditions—A comparative study of patterning methods based on PLL-g-PEG background passivation

Cited by 94 publications

References 41 publications

Locomotion of fish epidermal keratocytes on spatially selective adhesion patterns

Locomotion of fish epidermal keratocytes on spatially selective adhesion patterns

Micropatterning neuronal networks

Integration column: microwell arrays for mammalian cell culture

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